Compositions and methods for treating a pathogenic metabolic condition of the gut microbiota and derived diseases

ABSTRACT

The invention relates to the use as a drug or medical nutritional product in humans to treat a pathological dysbiosis and/or to prevent or treat at least one derived metabolic disease chosen from among excess weight with comorbidity, obesity, diabetes, heart and vascular diseases, atherosclerosis, degenerative bone disease, neurodegenerative disease, metabolism-related cancers, autoimmune diseases, steatosis, metabolic steatohepatitis, and chronic inflammatory diseases of the intestine, by increasing the  Christensenella  bacterial genus in the intestine, of a composition comprising a mixture of active ingredients consisting of at least:
         a whey hydrolysate with a molecular weight comprised between 200 and 10,000 Daltons,   a whey isolate and/or a concentrate, and   calcium caseinate.

CROSS-REFERENCE TO RELATED APPLICATION

This application is a continuation application of U.S. patentapplication Ser. No. 14/700,462 filed Apr. 30, 2015 which claims thebenefit of priority from French application 1552740, filed Mar. 31,2015, the entire disclosure of the applications is incorporated hereinby reference.

TECHNICAL FIELD OF THE INVENTION

The present invention relates to the use of a composition as a drug or amedical nutritional product for the treatment of a pathogenic medicalcondition of the gut microbiota caused by a pathological dysbiosis ofmetabolic origin and/or to prevent and treat diseases derived from thatpathogenic condition.

BACKGROUND OF THE INVENTION

The microbiota is a set of microorganisms (bacteria, archaea, virusesand eukaryotes) that are specific to each individual. Thesemicroorganisms are located on the skin, in the mouth and mainly in thedigestive system, which, for 10¹² cells per gram and since there are 1.5to 2 kg of microbiota, counts the presence of millions of differentspecies, i.e., between 3.3 and 8 million according to the experts, andbillions for all microorganisms. This microbiota contains more than onehundred fifty times the genes of the host genome.

The microorganisms of the gut microbiota are classified by kingdom (forexample, bacteria), phylum (for example, Firmicutes), class (forexample, Clostridia), order (for example, Clostridiales), family (forexample, Christensenellaceae), genus (for example, Christensenella) andspecies (for example, Christensenella minuta).

Each individual has his own microbiota, which comes from his history androots. However, in a cohort, 75% of species are found in 50% of thecohort and 57% in 90% of the cohort. More than 85% of species are sharedbetween Europe, the USA and Japan.

The gut microbiota is not homogenous. For a same individual, it variesin quantity and quality from the stomach (10¹ with lactobacillus,vellonella, helicobacter) to the duodenum, jejunum, ileum (10³ to 10⁷with bacilli, streptococcaceae, actinobacteria, actinomycinaeae,corynebacteriaeae) and lastly the colon (10¹² with lachnospiraceae,bacteriodetes). Certain bacteria (clostridium, lactobacillus,enterococcus) are also found in the mucus that coats the gut wall. Themajor phyla are relatively stable in an individual, and the differencesare found in terms of the species, often by several percentage points ofthe total microbiota. Thus, pathogenic conditions are difficult todetect, since they come from these specific species.

The anomalies of the host (genetic and environmental) lie in animbalanced pathogenic flora called pathogenic dysbiosis; the imbalancethreshold is difficult to determine, as are the variations in genusesand species, unless there is a biomarker for the anomaly or the disease.For several years, it has been known that the gut microbiota is not onlyinvolved in the digestion and transformation of food into energy, butthat it also plays a major role in maintaining good health and in theappearance of several diseases. Recent research suggests that thebehavior of the brain, the enteroendocrine system or neurovegetativesystem, or degenerations, are controlled by the microbiota. It is alsoknown that the gut microbiota is a major regulator in immunity andparticipates heavily in regulating gene expression of the host.

Through dysbiosis, i.e., a change in the composition of the microbiota,the microbiota may be associated with direct pathogenic metabolicdisorders such as type 2 diabetes or cardiovascular disease.Furthermore, certain components of the microbiota have been associatedwith other diseases such as degenerative diseases, gut diseases andcertain types of cancer.

The gut microbiota is also directly correlated with the host's weight.Excess weight with comorbidity and obesity, which are defined by a bodymass index (BMI) from 25 to 29.9 for overweight and over 30 for obesity,are in fact associated with a pathological dysbiosis of metabolic originof the gut microbiota.

Excess weight with comorbidities, obesity and metabolic diseases broadlyspeaking, are devastating our modern societies, in highly developedcountries, emerging countries or even developing countries.

The search for reasons for this epidemic has become critical so as notto see a drop in life expectancy in the next 25 years, and particularlyto avoid increasing the number of people in poor health and who aredependent in the end-of-life phase.

The key causes for weight gain are imbalanced food intake, in particularsaturated fats, fructose and carbohydrates, which may or may not beassociated with a highly sedentary lifestyle relative to food intake,i.e., relative to the quantity of energy. However, not everyone respondsin the same way to the same food intake. There are in fact in particulardifferences in terms of the absorption and storage of energy in the formof fats, and these differences are related to the pathological conditionof the gut microbiota.

In case of excess weight and obesity, it has been observed that:

-   -   the richness and diversity of the microbiota decreases    -   certain families, genuses and species of pathogens take        precedence over others    -   the pathogenic microbiota has the ability to extract more energy        from foods and non-digestible fibers.

Furthermore, in overweight and obese individuals, a calorie restrictionwith rebalancing of the diet increases the total richness of the floraby losing body fat mass, but there is a difference in results of thediet depending on the richness of the initial flora. For microbiota thatare initially depleted, the results are lower, in particular on thedecrease of inflammation and insulin resistance.

Furthermore, the analysis of the microbiota of thin or even anorexicpeople compared to the obese has established a correlation with certainmicroorganisms.

A correlation between the phyla and excess weight/obesity has beensearched for, and a relationship has been found between the 2 biggestphyla: the relative ratio of Firmicutes to Bacteriodetes increases withthe BMI, and the effect on weight loss results in a decrease in theratio (Damms-machado a 2015, Pekkala s 2015, Remely m 2015, eslinger aj2014, Bervoets l 2013, Fava f 2013), though this observation has alsobeen invalidated by results demonstrating the opposite(Rahat-Rozenbloom, 2014 Xu P2012, Sefcíková Z 2010).

It is known that the composition of an individual's microbiota isrelatively homogenous over time in terms of the phyla (aside fromtemporary incidents such as antibiotic treatment) but that the genusesmay vary, and it is also known that among several individuals, the phylavary depending on the age, geographical location, ethnicity, lifestyle,etc. It is therefore impossible to establish a standard microbiota forgood health, but it is possible to look for the genuses and species thatindicate a condition with or without pathology.

Certain enterotypes with the assembly of positive or negativerelationships have been determined. In particular, three enterotypes arepredominant throughout the world depending on the primacy of the mostsignificant phylum—bacteriodetes, provotella and ruminococcus—but nodirect correlation has been able to be established between theseenterotypes and metabolic diseases.

For the composition of the microbiota until 2012, it was accepted thatthe microbiota was built through outside influence and that it wascompletely acquired from 0 to 3 years through the environment and diet.In a study from November 2014, an international team run by Bruce Ley(Cornell University, NY) published, under the oversight of JuliaGoodrich, an article (“Human genetic shape the gut microbiome”, CellNovember 6, 159 789-799), which invalidates the single concept ofacquisition by introducing the notion of inheritability for severalfamilies, genuses and species; she made this observation by studying aBritish cohort of monozygotic and dizygotic twins and control persons.The strains of the gut microbiota demonstrated as being inheritable are:

-   -   families of bacteria in the Firmicutes: lachanospiraceae,        ruminococcaceae and christensenellaceae,    -   an archea in the Methanogens: M. Smithii.

The family of the Christensenellaceae is the most transmissible family.It in particular comprises the Christensenella genus, and in particularthe Christensanella minuta species, which has been defined andcultivated recently by Masami Morotomi in Tokyo, described in the study“Description of christensenella minuta gen. nov.sp.nov. isolated fromhuman faeces, which forms a distinct branch in the other clostridiales,and proposal of christensenellaceae” (M Morotomi 2012 inter. Jour. ofsystematic and evolution microbiology 62 144-149).

In children, at birth, the richness of Christensenellaceae is 20% in themeconium versus 8.6% for the mother. This proportion next decreasesrelatively (to be approximately 5%) through the rapid increase of theother phyla, in particular those that will become preponderant, such asFirmicutes, Bacteriodetes, Proteobacteria and Actinobacteria with theintroduction of food (for example, animal fat or proteins) and theenvironment.

The presence of Christensenella is associated with a low BMI as well asa “good health” condition. The higher the relative quantity ofChristensenella is, the more the host tends to be lean and healthy. Dueto its low relative presence, Christensenella would become the biomarkerfor pathogenic dysbiosis, a metabolic pathological condition inparticular leading to excess weight with comorbidity, obesity andmetabolic diseases.

The family of christensenellaceae is the leader of a “consortium” madeup of the following families, ranked by decreasing order of importancein metabolic action: dehalobacteriaceae (firmicutes), unclassifiedclostridiales, tenericutes unclassified geniuses ML615J-28 and RF39,methanobacteriaceae (euryarchaeota), firmicutes unclassified genusSHA-98, peptococcaceae, verrucomicrobiaceae.

The Christensenellaceae family is often positively linked to theabundance of Methagenes, which in turn is correlated to the productionof butyrate and propionate, but Christensenellacea does not need thislink to express itself in the metabolism, since it contributes directlyto the phenotype of the host with which it is associated.

Julia Goodrich (“Human genetic shape the gut microbiome” Cell November6, 159 789-799) continued her observation in germ-free mice seeded withobese microbiota in 2 groups, only one of which received an implantationof Christensenella, and that presence caused weight gain of less than30%.

The Christensenellaceae genus is therefore recognized as a marker ofleanness, when it is present in sufficient quantities in the gutmicrobiota. Furthermore, it is a marker of excess weight and obesitywhen it is present in small quantities in the gut microbiota. Therelative increase of Christensenellacea guarantees weight loss, inparticular on the fat mass, and is a driving force for metabolicimprovement.

Furthermore, these Christensenellacea bacteria would be involved inother metabolic diseases such as diabetes, heart and vascular disease,atherosclerosis, degenerative bone diseases, neurodegenerative diseases,cancers related to the metabolism, autoimmune diseases and metabolicsteatosis and steatohepatitis.

Currently, there is a need for a product capable of acting on the gutmicrobiota to be able to be used to treat the pathogenic microbiota, andconsequently to prevent and treat derived metabolic diseases, such asexcess weight with comorbidity, obesity, etc.

SUMMARY OF THE INVENTION

To meet this need, the present invention proposes to use a compositioncapable of increasing the relative quantity of Christensenellacea in thegut microbiota.

To that end, the invention relates to the use of a compositioncomprising a mixture of active ingredients made up of at least:

-   -   a whey hydrolysate with a molecular weight comprised between 200        and 10,000 Daltons,    -   a whey isolate and/or a concentrate, and    -   calcium caseinate,        to be used as a drug or medical nutritional product in humans to        treat a pathological condition of the microbiota, namely a        metabolic pathological dysbiosis characterized by a drop in        Christensenellacea in the gut microbiota. The composition may        also be used for the prevention or treatment of at least one        derived metabolic disease, in particular a metabolic disease        marked by a deficiency of Christensenellacea in the gut        microbiota, in particular a metabolic disease marked by a        deficiency of Christensenellacea in the gut microbiota chosen        from among excess weight with comorbidity, obesity, diabetes,        heart and vascular diseases, atherosclerosis, degenerative bone        diseases, neurodegenerative diseases, metabolism-related        cancers, autoimmune diseases, steatosis, metabolic        steatohepatitis and chronic inflammatory diseases of the        intestine. The composition treats the pathological microbiota in        particular by causing a relative increase in the intestine of        the Christensenella bacterial genus (phylum Firmicutes, class        Clostridia, order Clostridiales, family Christensenellaceae,        genus Christensenella).

Advantageously, such a composition is able to bring about a relativeincrease in the quantity of Christensenella in the gut microbiota so asto combat metabolic pathogenesis. It may also be used as a drug ormedical nutritional product to rebalance the gut microbiota byincreasing the Christensenella genus and to prevent and combat diseasesresulting from that pathological imbalance of the gut microbiota.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1: a diagram of the results of table 1 relative to the relativeabundance of Christensenella,

FIG. 2: a barcode table, done by E. Prifti, of the results obtained inthe study done on the gut microbiota of patients treated with a placeboand patients treated according to the invention.

DETAILED DESCRIPTION

The invention relates to a composition comprising a mixture of activeingredients made up of at least:

-   -   a whey hydrolysate with a molecular weight comprised between 200        and 10,000 Daltons,    -   a whey isolate and/or a concentrate, and    -   calcium caseinate,        to be used as a drug or medical nutritional product in humans to        treat a pathological dysbiosis characterized by a deficiency of        Christensenella in the gut microbiota and/or to prevent or treat        at least one metabolic disease derived from that pathological        condition of the gut microbiota chosen from among excess weight        with comorbidity, obesity, diabetes, heart and vascular        diseases, atherosclerosis, degenerative bone disease,        neurodegenerative disease, metabolism-related cancers,        autoimmune diseases, steatosis, metabolic steatohepatitis, and        chronic inflammatory diseases of the intestine.

The composition acts through a relative increase in the intestine of theChristensenella bacterial genus.

The composition is known, but surprisingly and unexpectedly, it acts onthe microbiota, and in particular is capable of increasingChristensenella.

Within the meaning of the invention, “deficiency” of Christensenella inthe gut micrograft refers to a level of Christensenella in the gutmicrograft representing less than 0.01% of the total bacterial genusesdetected in the microbiota of a given individual (collected in thestool, the abundance being measured preferably using a Fish sequencingtest, or qPCR or meta-analysis).

“Christensenella” refers to bacteria in phylum Firmicutes, classClostridia, order Clostridiales, family Christensenellaceae, and genusChristensenella.

Within the meaning of the invention, “medical nutritional composition”or “medical nutritional product” or “medical food” or medical food forspecial medical purposes or dietary food for special medical purposesrefers to a food for therapeutic prevention or treatment use, used aloneor in combination with other therapies. It involves a nutritionalcompound, responding to a particular clinical situation, able toconstitute all or part of the food for the patients for whom it isintended.

Within the meaning of the invention, “whey concentrate” refers to a wheyextract obtained by concentrating a whey.

Within the meaning of the invention, “pathological dysbiosis” refers toa pathological condition characterized by an imbalance in thedistribution of the bacteria of the gut microbiota in terms of thephyla, classes, orders, families, genuses or species, leading to a riskof other metabolic diseases.

Within the meaning of the invention, “whey hydrolysate” refers to anymolecule or mixture of molecules obtained using the method comprising achemical hydrolysis or enzyme hydrolysis step for the whey.

Within the meaning of the invention, “whey isolate” refers to a wheyextract that contains less than 1% lactose and fats.

Within the meaning of the invention, “drug” refers to a product havingobtained a marketing authorization as a treatment product for a disease.

Within the meaning of the invention, “microbiota” refers to the gutmicrobiota.

The whey hydrolysate of the composition according to the invention has amolecular weight comprised between 200 and 10,000 Daltons, preferablybetween 200 and 3,500 Daltons. It is essentially made up of di- andtri-peptides.

Preferably, it is a peptide whey hydrolysate comprising at least 90%peptides by dry weight of the hydrolysate.

The whey isolate and/or concentrate preferably has a molecular weightcomprised between 15,000 and 20,000 Daltons.

The whey isolate is preferably manufactured from fresh milk and cheesedairies that do not pasteurize the milk to avoid the destruction of thebeta-lactoglobulin and alpha-lactalbumin, and which extract the whey byultrafiltration or microfiltration (filter size of 0.1 μm). The isolateobtained by ion exchange is less suitable due to its lowbeta-lactoglobulin and alpha-lactalbumin content level. The isolatecontains less than 1% lactose and fats, and its peptide concentration ispreferably at least 90% by dry weight.

The whey concentrate is preferably obtained from a cheese dairy wheywithout pasteurization, containing beta-lactoglobulin, alpha-lactalbuminand glycomacropeptides. The peptide concentration of the concentrate ispreferably at least 80% by dry weight.

Furthermore, the calcium caseinate used in the composition according tothe invention preferably has a molecular weight comprised between 20,000and 35,000 Daltons.

Preferably, the weight ratio between the calcium caseinate and themixture formed by the hydrolysate and the whey isolate and/orconcentrate is from 0.8 to 1.2 in the composition. According to oneparticularly suitable embodiment, the mixture of active ingredients ofthe composition also comprises a mixture of amino acids.

The amino acids present in composition are preferably at leasttryptophan, glutamine, leucine, arginine and/or taurine, but thecomposition may contain other amino acids, such as isoleucine, valine,phenylalanine or threonine. Very preferably, the composition accordingto the invention comprises at least tryptophan, leucine, arginine andtaurine.

When tryptophan is present, it must make up between 6 and 9%, preferablyapproximately 7%, by weight of the neutral amino acids present in thecomposition (leucine, isoleucine, valine, phenylalanine, tyrosine andtryptophan).

When arginine and taurine are present, the weight ratio of arginine totaurine is comprised between 1.5 and 2.

In addition to the mixture of hydrolysate, whey isolate and/orconcentrate and calcium caseinate, and amino acids, the mixture ofactive ingredients of the composition according to the invention maycomprise one or more elements chosen from milk calcium, magnesium,vitamin B6, vitamin B9, vitamin E, vitamin D, zinc and chromium.

Likewise, the composition may contain essential fatty acids, inparticular omega-3s. Preferably, these are omega-3s of plant origin,with a high proportion of EPA.

According to one preferred embodiment, the mixture of active ingredientsof the composition according to the invention comprises at least

-   -   8 to 12% whey hydrolysate,    -   15 to 20% whey isolate and/or concentrate,    -   20 to 25% calcium caseinate,        the percentages being given by dry weight of all of the active        ingredients present in the composition (excluding any        excipients).

The composition may also contain freely added elements, such as aminoacids, vitamins and minerals, which are added to the native componentsof the whey hydrolysate, the whey isolate, the whey concentrate and thecalcium caseinate.

The composition according to the invention is preferably made up of atleast:

-   -   1.5 to 3% tryptophan,    -   12 to 20% branched amino acids,    -   6 to 10% aromatic amino acids,    -   0.8 to 1.5% taurine,    -   1.6 to 3% arginine,    -   1.2 to 3% milk calcium,    -   0.5 to 1% magnesium,    -   0.4 to 1% omega-3s,    -   1 to 2 mg of vitamin B6 per 50 g of composition without        excipients,    -   5 to 15 mg of zinc per 50 g of composition without excipients,    -   1 to 3 μg of vitamin D per 50 g of composition without        excipients,    -   75 to 150 μg of chromium per 50 g of composition without        excipients,    -   100 μg of vitamin B9 per 50 g of composition without excipients,    -   10 mg of vitamin E per 50 g of composition without excipients,        the percentages being given by dry weight of all of the active        ingredients present in the composition (excluding excipients),        part of the components coming from the whey hydrolysate, the        whey isolate, the whey concentrate and the calcium caseinate,        and the rest being added freely in the form of amino acids,        vitamins and minerals.

The branched amino acids of the composition are made up of leucine,isoleucine and valine, preferably:

-   -   50 to 60% leucine,    -   18 to 25% isoleucine, and    -   20 to 28% valine,        and the aromatic amino acids of tryptophan, phenylalanine and        tyrosine, preferably:    -   15 to 24% tryptophan,    -   38 to 46% phenylalanine, and    -   35 to 43% tyrosine.

The composition according to the invention may be obtained using amethod as described below:

-   -   a first mixture is obtained by mixing components in the        following order: calcium caseinate, whey isolate, whey        concentrate, whey hydrolysate, free amino acids, magnesium and        milk calcium. The pH must be around 7 and stabilized at that        level.    -   adding vitamins, minerals and fatty acids to the initial        mixture.

A powder is thus obtained that can be turned into a tablet or liquid, orused in its powdered form in pouches, sticks, cans or gel caps, forexample.

When the composition according to the invention is administered orallyin a sufficient quantity, it makes it possible to modify the compositionof the gut microbiota by causing a relative increase in the quantity ofChristensenella in the microbiota. The composition makes it possible toreestablish the metabolic functionality through a relative increase ofChristensenella within the general population of the microbiota, or atleast a level making up 0.01% of the total of the bacterial genusesdetected in the microbiota of a given individual (collected in thestool, abundance preferably being measured using a Fish sequencing test,or qPCR or meta-analysis).

This increase makes it possible to combat metabolic pathologicaldysbiosis caused by a deficiency of Christensenella in the gutmicrobiota.

The increased relative quantity of Christensenella also makes itpossible to prevent or combat metabolic diseases originating from ametabolic pathological dysbiosis characterized by a relative deficiencyof Christensenella in the gut microbiota, and in particular:

-   -   excess weight with comorbidity    -   obesity    -   diabetes (and pre-diabetes)    -   heart and vascular diseases    -   atherosclerosis    -   degenerative bone diseases    -   neurodegenerative diseases, in particular Parkinson's disease        and Alzheimer's disease    -   cancers related to metabolism, in particular esophageal, colon,        endometrial, kidney and breast cancer for postmenopausal women,    -   autoimmune diseases,    -   steatosis, metabolic steatohepatitis and NASH (nonalcoholic        steatohepatitis),    -   chronic inflammatory diseases of the intestine.

The composition according to the invention is particularly useful forpeople with a relative proportion of Christensenella below 0.01% of allbacterial genuses detected in the microbiota (collected in the stool,abundance preferably being measured using a Fish sequencing test, orqPCR or meta-analysis), and for people with a relative proportion ofChristensenella minuta in the microbiota below 0.005% of the totalabundance of the gut flora, i.e., the total bacterial species detectedin the microbiota (collected in the stool, abundance preferably beingmeasured by a Fish sequencing test, or qPCR or meta-analysis). Differentmethods may be used to analyze the microbiota (the collection generallybeing done in the stool):

-   -   culture using traditional bacterial culture methods known by        those skilled in the art    -   biological methods through the 16SrRNA marker    -   sequencing either using the SANGER method with Operational        Taxonomic Units, or by pyrosequencing at the 16SrRNA (method        used in studying the composition)    -   imprint by electrophoresis, polymorphism and ribosomal,    -   DNA with micro areas,    -   FISH (fluorescence in situ hybridization) and qPCR (quantitative        polymerase chain reaction) with amplification of a group,    -   meta-analysis with meta-genomics (composition and function of        all of the genes), metaproteomics (protein analysis),        metabolomics (metabolic profiling) and metatranscriptomics        (through the RNA).

According to one alternative, in case of an extreme medical emergency,it is possible, for people with a Christensenella deficiency, to implantbacteria of the Christensenella genus in the intestine prior to theadministration of the composition according to the invention, by using aspecific probiotic for that type of bacteria in a capsule or by directimplantation by lead in the jejunum. The implantation should represented75 to 100 mg of Christensenella bacteria. The implanted bacteria may bebacteria obtained by culturing bacteria from the genus Christensenellataken from the stool of the treated person if there are traces or frombacteria obtained by culturing bacteria from the genus Christensenellataken from the stool of lean and healthy people.

The use of the composition according to the invention in appropriatequantities makes it possible, optionally with prior implantation, tobring the presence of Christensenella to a minimum relative proportionof 0.01% relative to all of the taxa of the microbiota collected in thestool. The composition may also make it possible to obtain a relativequantity of Christensenella minuta (when the analysis method makes itpossible to go down to the species level) of at least 0.0075% (relativeto all of the taxa of the microbiota collected in the stool).

Surprisingly, the composition according to the invention is also capableof regulating, in the microbiota, the families of bacteria known fortheir relatively significant presence in excess weight, obesity andmetabolic diseases, chosen from among clostridiales, in particularLachnospiraceae and clostridiaceae.

Also astonishingly, it is also able to regulate, in the microbiota, thefamilies of bacteria known for their ability to increase the quantity ofenergy taken from food, chosen from among bacteria which yield butyrateand propionate by fermentation, but also increase energy through thedigestion of fibers that are not normally digested, namely: Roseburia,Faecalbacterium (in particular the species prausnitzii), Akkermansia (inparticular the species municiphali), Eubacterium (in particular thespecies rectale), Methanobrevibacter (in particular the speciessmithii). This additional energy transformation is one of the likeliestcauses of the excess weight epidemic and metabolic dysfunction. It isnecessary to decrease these “energizers”, but without creating adeficiency, since the microbiota is a major consumer of butyrate,propionate and acetate (in order of importance); the genus Akkermansiaand species Akkermansia municiphali (also inheritable) are particularlyimportant, since they are the main producers of propionate, whichguarantees proper operation of the gut barrier by limiting the passageof LPS; in fact, the latter, induced by diets too high in fat (saturatedin particular), is the first to blame for metabolic dysfunction,steatosis, hepatitis and the formation of adipose tissue. The regulationby the invention of 2 inheritable genes christensenella and akkermansiathat are difficult to alter is essential to restore the proper operationof the metabolism and also serve as markers for success of theintervention using treatment according to the invention.

The composition according to the invention may assume any suitable formfor oral administration. It may in particular assume the form of apowder or granules, ready-to-use beverages, bars or extruded products,capsules, gel caps, dispersible tablets or any other suitablepharmaceutical form, the composition being supplemented with excipientsand traditional fillers known by those skilled in the art.

The daily dose of composition according to the invention (dose ofmixture of active ingredients without the excipients) is preferablycomprised between 66 and 110 g, preferably in two administrations of 33to 55 g distributed one in the morning as breakfast or at 11 AM as asnack, and the other as an afternoon snack.

Advantageously, with the goal of weight loss, the bioavailability in thebody of the amino acids, peptides and proteins present in thecomposition in the body is comprised between 10 minutes and 5 hours,which allows a rapid action that lasts over time so as to limit thequantity of food to be consumed daily.

Furthermore, the presence of the milk calcium makes it possible toimprove the palatability of the dietary product according to theinvention, in particular by hiding the bitter taste of the wheyhydrolysate, such that it contributes to eliminating the risk of peopleceasing to consume it for taste reasons and abandoning their treatmentbefore it is complete.

The composition according to the invention falls within customizedmedicine and is a major advance in particular in pharmaco-metabolicmedicine.

The composition according to the invention may in fact be used as atreatment adapted to the metagenomics of the treated individual.

Thus, before treatment using the composition according to the invention,a test may be done to detect the presence or absence of Christensenellaand its proportion in the microbiota. This test makes it possible toestablish a likelihood of total or partial success or failure of thetreatment. The test may be repeated at the end of treatment to analyzethe increased level of the bacteria, then at one year to measure thelastingness of the treatment and risk of relapse with the possibility ofperforming another preventive treatment. The test may also relate toother bacteria, in particular Akkermansia. The advantage lies in nolonger treating blindly and monitoring the patient's chronic disease forlife.

The invention thus makes it possible to resolve the problems oftherapeutic failures, of which there are many, either due to abandonmentof the treatment in progress, and most often at the beginning, or due toregaining weight after several months or years after significant weightloss. Failures are attributed to the lack of motivation or disinterestin making permanent efforts, but the blockage may be due to the near orcomplete absence of Christensenella in the microbiota.

The invention is now illustrated through a non-limiting example of adietary composition, assuming the form of a powder of 55 g (activeingredients and excipients) packaged in a pouch.

This composition is obtained from the following active ingredients:

-   -   5 g of whey hydrolysate with a molecular weight between 200 and        3,500 Daltons,    -   10 g of whey isolate and/or concentrate with a molecular weight        between 15,000 and 20,000 Daltons,    -   13 g of calcium caseinate with a molecular weight between 20,000        and 35,000 Daltons,    -   1 mg of vitamin B6,    -   10 mg of zinc,    -   0.45 g of taurine,    -   40 μg of chromium,    -   10 mg of vitamin E,    -   100 μg of vitamin B9,    -   2.5 μg of vitamin D,    -   270 mg of plant omega-3s,    -   q.s. 4.2 g of leucine,    -   q.s. 0.8 g of tryptophan,    -   q.s. 0.9 g of arginine,    -   q.s. 0.75 g of milk calcium,    -   q.s. 0.36 g of magnesium.        “q.s. X g” of an element of the competition refers to the total        quantity of that element in the composition: quantity provided        by the proteins (calcium caseinate, whey isolate, whey        concentrate, whey hydrolysate) and completed by the addition of        the element in free form to arrive at X g.

The invention is now illustrated through a study showing the effect ofthe composition in the gut microbiota, in particular on Christensenella.

The study pertained to 54 subjects. This is an ancillary study to arandom, blind international clinical study with another active productas control. Two ancillary studies were conducted on 54 subjects, one onthe microbiota and the other on the adipose tissue.

The clinical study pertained to 108 patients whose characteristic wasbeing overweight or obese with at least 3 proven comorbidity factors;the subjects were enrolled during a visit comprising the morphologicalcriteria, a CT scan, a DEXA, and a complete biological analysis. For theancillary study on 54 subjects, a sample of subcutaneous adipose tissueand stool samples were also taken. The subjects received either thecomposition according to the invention or an isocaloric active controlcorresponding to the composition according to the invention, but inwhich the whey was replaced by a pea protein, known for its action onthe microbiota.

The study of the microbiota included a withdrawal upon enrollment andanother during the visit in week 12, at the end of treatment with theproduct with moderate calorie restriction. The stool was frozen, and atthe end of the study, it was analyzed by pyrosequencing. The analyseswere conducted by the primary investigator, the Institut duCardiométabolisme et Nutrition de Paris.

Out of the 54 subjects, 27 were treated with the composition accordingto the invention (composition of the example) and 27 with the controlproduct.

Stools were collected upon enrollment and at the end of treatment inweek 12 with the 2 products and a calorie restriction of 600 kcal.

The results for the study of the microbiota are shown in table 1 below,and in FIGS. 1 and 2.

For the results shown in FIG. 2, the following items should be noted.

-   -   the Wilcoxon test between genuses was done based on a normalized        abundance,    -   at value p<0.05, 16 genuses are more abundant with placebo        treatment and 26 genuses with the treatment according to the        invention,    -   only one genus resists multiple tests, i.e., Christensenella.

TABLE 1 STANDARD Artificial bacteria community (1) INVENTION Relative PRPR READS READS presence READS W 0 W 12 W 0 W 12 POPULATION 27 TOTAL TAXA100 82394 100 100 75,875 74,220 FIRMICUTES 72.9 60019 67.2 68.2 51,26050,633 BACTERIODETES 20.7 17057 19.9 22.4 14,863 16,660 PROTEOBACTERIA3.0 2470 7 4.8 5,430 3,538 ACTINOBACTERIA 5.0 4148 2.3 1.7 1,721 1,264FIRMICUTES/ 3.5 N 3.4 3.0 BACTERIODETES OVERWEIGHT 22 18,147 22.2 19.817,093 14,670 CONSORTIUM (2) ENERGIZER 5.2 4,290 8.7 8.2 6,592 6073CONSORTIUM (3) of which NC NC 0.0161 0.0337 12 25 AKKERMANSIACHRISTENSENELLACEAE 11.5 9,494 11.4 11.4 8,389 8,486 CONSORTIUM (4)CHRISTENSENELLA 0.0097 8 0.0028 0.013 2 10 GENUS CONTROL INVENTION PR PRREADS READS Δ % W 0 W 12 W 0 W 12 Δ % POPULATION TOTAL TAXA −1.5 100 10073,838 72,365 1.2 FIRMICUTES 12.0 69.3 63.5 51,953 45,953 7.4BACTERIODETES 16 20.5 25.9 15,063 18,724 21.3 PROTEOBACTERIA −20.6 5.86.8 4,281 4,962 16.5 ACTINOBACTERIA −16.3 2.6 2.1 1,896 1,496 −12.3FIRMICUTES/ 3.4 2.5 NS BACTERIODETES OVERWEIGHT −9 25.3 25.0 18,73018,110 0.5 CONSORTIUM (2) ENERGIZER −6 10.5 12.0 7,801 8,670 13CONSORTIUM (3) of which 118 0.0097 0.0186 7 13.5 57% AKKERMANSIACHRISTENSENELLACEAE 1.6 0.07 11.0 8,982 8,630 −3.8 CONSORTIUM (4)CHRISTENSENELLA 380 0.002 0.0027 1.5 2 60% GENUS (1) there is no idealmicrobiota model, since it is individual and depends on the genetics andexperience of each person; for the study, a standard of the cultivatedfamilies was developed; this standard was used to replace the results ina context (2) the overweight consortium comprises many families, inparticular in the clostridiales order, but 2 appear to be important dueto their number and force of impact: lachnospiraceae and clostridiaceae(3) the consortium of “energizers” are the bacteria which yield butyrateand propionate by fermentation, but also increase energy through thedigestion of fibers not normally digested; it is made up of: Roseburia,Faecalbacterium, Akkermansia, Eubacterium, Methanobrevibacter. Balanceis particularly important in this group. (4) the christensenellaceaeconsortium comprises the families: Christensenellaceae,Methanobacteriaceae, Dehalobacteriaceae, Peptococcaceae,Verrucomicrobiaceae, Clostridiales-nonclassified (in the present study,unclassified clostridiales were included, as well as others to giveweight to that family).

-   -   Out of the total microbiota, a relative depletion should be        noted for both the Invention and Control groups relative to the        standard, which is explained by the metabolic situation of those        2 groups; treatment does not improve this general situation,        which is logical, since 12 weeks is an insufficient length of        time for the major phyla.    -   Out of the mass of traditional major firmicute and bacteriodete        phyla and their relationship, there is a relative depletion, but        above all for the firmicutes, which can be explained by the        insulin resistance and very advanced inflammation situation; the        situation improves slightly on average for groups treated with        the composition according to the invention. In terms of the        relationship (currently controversial), the situation is        improving, but more for the control group also through a        deterioration of the firmicutes, which shows that this ratio is        not particularly interesting.    -   In the other 2 major phyla, the Proteobacteria are high and the        Actinobacteria are low; these results perfectly reflect the        metabolic situation of these subjects; there is a favorable        relative evolution for the group according to the invention in        both phyla.    -   The overweight consortium evolved favorably for the invention,        but in the opposite direction for the control.    -   The energizer consortium, which is high in both groups, is being        resized for the group according to the invention, unlike the        control group; Akkermansia, which was particularly low, is        resized in both groups, but with a more significant increase for        the group according to the invention.    -   The Christensenellaceae consortium (Goodrich 2013) increases in        the groups treated according to the invention, reflecting the        improvement in terms of the metabolic level and weight loss.    -   In the study of the Christensenellaceae family, the unclassified        genuses not belonging to Christensenellacea are crucial (>99%).        According to a 2013 publication by Rajilic-Strojanovic, these        unclassified genuses are involved in the inflammation of        inflammatory bowel disease. Here there is a relative decrease        for treatment according to the invention from 1.26 to 1.12 and        an increase for the control from 0.83 to 0.89. However, since        several genuses should exist in this unclassified part and that        could evolve in divergent ways, it is impossible to draw        conclusions from this; it is advisable to wait for new        discoveries in these unclassified genuses.    -   The analysis of the christensenella genus is at the core of this        study on microbiota. Christensenellaceae is an “inheritable”        bacteria and is not acquired and not sensitive to diet; in this        category, it seems to be the most important in the group. This        implies that for people with a low Christensenellaceae level, it        is “vital” to reestablish a healthy metabolism, to increase this        genus rather than all of the microbiota with prebiotics or        probiotics, which can cause increases in undesirable classes,        such as methanogens or helicobacters. The invention has a        targeted action on the bacteria, useful for the metabolism. The        regulation of Akkermansia is second in terms of priority, then        those of the consortium on weight and energizers.

Furthermore, Christensenellaceae is a marker bacteria for “normal”weight from the metabolic perspective, and a good health marker,therefore a microbiota that works regularly without pathogenicity bothgenetically and in terms of lifestyle. Furthermore, Goodrich hasdemonstrated that the implantation of Christensenellaceae reversed theweight curve through a lower weight gain in mice, which is reflected inhumans by weight loss.

The action of Christensenellaceae is particularly essential formetabolic regulation in order to erase the pathogenic factors thatresult in metabolic diseases.

The Christensenellaceae genus is relatively underrepresented in bothgroups (0.0028 and 0.002 versus 0.0097) relative to the standard, whichconfirms the decrease of this genus in a population suffering frommetabolic issues. The action of the 2 products causesChristensenellaceae to increase from 0.0028% to 0.014% for the groupaccording to the invention (380%), with a significance relative to theinitial level (p=0.002), while the control goes from 0.002% to 0.0028%(+60% and not significant). In 12 weeks, the group treated according tothe invention very clearly goes above the standard (0.014% versus0.0097%), and it is significant relative to the control (0.004).

Furthermore, it should be noted that of all of the genuses comprised inthe human microbiota (1,500 known at this time), the only significantgenus in the study on the invention that emerges from all of thestatistical tests is Christensenellaceae, which appears to be one of themost significant genuses for metabolic pathological conditions of themicrobiota and derived metabolic diseases.

Furthermore, the clinical results and the ancillary study on the adiposetissue are shown in table 2 below:

TABLE 2 Composition according to the invention TOTAL FAS TOTAL FAS ΔBase Line % AT 12 WEEKS CLINICAL STUDY POPULATION 108 VISCERAL BODY FATMASS CT 192.3 −10.4%  Scan cm² TOTAL BODY FAT MASS CT 489.7 −8.9% Scancm² BODY FAT MASS DXA kg 37.0 −8.9% LEAN BODY MASS DXA kg 51.2 −0.1%WAIST SIZE cm 104.5 −4.2% WEIGHT kg 92 −3.9% TAS mm 131.7 −2.7% TAD mm81.6 −4.0% GLYCEMIA mmol 5.5 −2.8% INSULINEMIA mUI/l 12.6 −19.0%  HOMAIR 3.1 −20.5%  HOMA B 124 −9.3% HOMA S 59.9 +40.6%  TOTAL CHOLESTEROLmmol 5.2 −7.2% HDL mmol 1.2 +0.2% LDL mmol 3.2 −7.6% TRIGLYCERIDES mmol1.6 −9.6% TNF pg/ml 1.9 −10.2%  ANCILLARY STUDY ADIPOSE TISSUE GLYCEROLpg/ml 16 −20.1%  IL6 3707 −13.3%  0² ENERGY CONSUMPTION μM 0.002  +63%ADIPONECTIN pg/ml 11426 +13.3% 

One can see that the composition used according to the invention has anunquestionable action on the metabolic function of the adipocyte, whichresults in increased adiponectin in the adipose tissue, which reflectsthe two basic dysfunctions at the origin of many metabolic anddegenerative diseases:

-   -   inflammation with a decrease in IL6 in the adipose tissue and        TNFα in the plasma, which is explained by the activation of p105        on the adipocytes, which regulates NFκB, the inflammatory        cytokine modulator.    -   insulin resistance, which improves with a decrease in the        insulinemia, decreased HOMA IR, but above all a very significant        improvement in insulin sensitivity by HOMA S.    -   two other functions on the adipose tissue should be noted: the        regulation of the lipolysis of the adipocytes of people who are        overweight (glycerol) and increased energy production (O²        consumption)    -   improved comorbidity factors: blood pressure, glycemia,        insulinemia, cholesterol, LDL and triglycerides, is both a        consequence of metabolic improvement and the direct action of        the active compounds of the invention, in particular amino acids    -   lastly, furthermore, no serious side effects were noted during        treatment aside from several temporary digestive difficulties.

According to the invention, it is thus possible to act on a pathologicaldysbiosis of the gut flora through a relative increase in theChristensenella genus, and consequently also to act on the metabolicdiseases marked by this pathology. The actions on other families,genuses or species reinforce the action of Christensenella.

What is claimed is:
 1. A method of treating a pathological dysbiosis ofthe gut microbiota characterized by a Christensenella genus deficiencyin the gut microbiota in an overweight or obesity patient and/or in apatient with metabolic syndrome, the method comprising administering tothe patient a composition comprising a mixture of active ingredientsmade up of at least a whey hydrolysate with a molecular weight comprisedbetween 200 and 10 000 Daltons, a whey isolate and/or a wheyconcentrate, and calcium caseinate.
 2. The method of claim 1, whereinthe patient is the patient whose Christensenella genus represents lessthan 0.01% of the total bacterial genuses detected in the gutmicrobiota.
 3. The method of claim 1, wherein administering thecomposition increases the Christensenella bacterial species in the gutmicrobiota of the patient and decrease permeability of the gut barrierand increase mucus.
 4. The method of claim 3, wherein the composition isadministered to the patient and the patient is monitored for increase inthe Christensenella genus greater than 0.01% of the total bacterialgenuses detected in the gut microbiota of the patient.
 5. The method ofclaim 1, wherein administering the composition increases theChristensenella minuta bacterial genus in the gut microbiota of thepatient such that the genus represents a proportion greater than 0.0075%of the total gut flora.
 6. The method of claim 1, wherein administeringthe composition increases the genus Akkermansia and species Akkermansiaprausnitzii that improve gut barrier plasticity.
 7. The method of claim1, wherein administering the composition regulates the bacteria knownfor their relatively significant presence in excess weight, obesity andmetabolic diseases, chosen from clostridiales and verrucomicrobialesorder in the microbiota of the patient and regulates the bacteria knownfor their ability to increase the quantity of energy taken from food,chosen from the Roseburia, Faecalibacterium, Akkermansia, Eubacterium,Methanobrevibacter genuses in the microbiota of the patient.
 8. Themethod of claim 1, wherein the patient is selected from a group ofpatients with a Christensenella deficiency and in whom bacteria of genusChristensenella was previously administered to the patient's intestineby at least one of the following methods: as a specific probiotic of thebacteria in a capsule; by direct implantation by lead; as bacteriaChristensenella obtained by culturing bacteria isolated from at leastone of the following: the intestine of the patient and isolated from theintestine of lean and healthy people or obese people withcardiometabolic syndrome.
 9. The method of claim 1, wherein the patientis also afflicted with at least one of the following diseases:Alzheimer's disease, Parkinson's disease and esophageal, colon,endometrial, kidney and postmenopausal breast cancer.
 10. The method ofclaim 1, wherein the whey hydrolysate has a molecular weight from 200 to3,500 Daltons, the whey isolate and/or the whey concentrate has amolecular weight in the range from 15,000 to 20,000 Daltons, the calciumcaseinate has a molecular weight in the range from 20,000 to 35,000Daltons.
 11. The method of claim 1, wherein the weight ratio between thecalcium caseinate and the mixture formed by the hydrolysate and the wheyisolate and/or concentrate is in the range from 0.8 to 1.2 and whereinthe whey hydrolysate comprises between 8% and 12%, the whey isolateand/or concentrate comprises between 15% and 20%, and the calciumcaseinate comprises between 20 and 25%, the percentages being given bydry weight of all of the active ingredients present in the composition.12. The method of claim 1, wherein the mixture of active ingredientsalso comprises a mixture of free amino acids selected from the groupconsisting of tryptophan, glutamine, leucine, arginine and taurine. 13.The method of claim 12, wherein the tryptophan makes up between 6 and 9%by weight of the neutral amino acids present in the composition.
 14. Themethod of claim 12, wherein the weight ratio of arginine to taurine isin the range from 1.5 to
 2. 15. The method of claim 12, wherein themixture of active ingredients further comprises one or more elementsselected from the group consisting of milk calcium, magnesium, vitaminB6, vitamin B9, vitamin E, vitamin D, zinc, chromium and omega-3s. 16.The method of claim 1, wherein the composition further comprises atleast the following: 1.5 to 3% tryptophan, 12 to 20% branched aminoacids, 6 to 10% aromatic amino acids, 0.8 to 1.5% taurine, 1.6 to 3%arginine, 1.2 to 3% milk calcium, 0.5 to 1% magnesium, 0.4 to 1%omega-3s, 1 to 2 mg of vitamin B6 per 50 g of composition withoutexcipients, 5 to 15 mg of zinc per 50 g of composition withoutexcipients, 1 to 3 μg of vitamin D per 50 g of composition withoutexcipients, 75 to 150 μg of chromium per 50 g of composition withoutexcipients, 100 μg of vitamin B9 per 50 g of composition withoutexcipients, 10 mg of vitamin E per 50 g of composition withoutexcipients, the percentages being given by dry weight of all of theactive ingredients present in the composition.
 17. The method of claim1, wherein the composition is administered in a form selected from thegroup consisting of powder, granules, a ready-to-use beverage, food bar,an extruded form, capsules, gel caps, and dispersible tablets.